摘要
目的:探讨FOXO3A对食管鳞状细胞癌(ESCC)细胞增殖、迁移、侵袭能力和自噬水平的影响。方法:采用RT-qPCR和Western blot法检测人正常食管上皮细胞HEEC、低分化ESCC细胞KYSE150和高分化ESCC细胞EC109中FOXO3A mRNA和蛋白的表达。将KYSE150或EC109细胞分别转染阴性对照siRNA(si-NC组)和靶向FOXO3A的siRNA(siFOXO3A组)。采用CCK-8实验检测转染72 h后细胞增殖活性,克隆形成实验评估细胞克隆形成能力,Transwell小室实验检测转染24 h后细胞的迁移和侵袭能力,Western blot法检测上皮间充质转化相关蛋白E-cadherin、N-cadherin和Vimentin的表达,免疫荧光法检测细胞中LC3B及p62的聚集度。结果:两种ESCC细胞中FOXO3A mRNA及蛋白表达水平均高于HEEC(P<0.05)。与si-NC组比较,siFOXO3A组细胞增殖活性、克隆形成数、迁移细胞数、侵袭细胞数均降低(P<0.05);N-cadherin、Vimentin蛋白表达降低,E-cadherin蛋白表达升高(P<0.05);p62聚集程度减弱,LC3B聚集程度增强(P<0.05)。结论:敲低FOXO3A可抑制ESCC细胞的增殖、迁移和侵袭能力,增强其自噬水平,发挥抑癌作用。
Aim:To investigate the effects of FOXO3A on the proliferation,migration,invasion,and autophagy of esophageal squamous cell carcinoma(ESCC)cells.Methods:RT-qPCR and Western blot were used to detect the expressions of FOXO3A mRNA and protein in human normal esophageal epithelial cell HEEC,poorly differentiated ESCC cell KYSE150,and well-differentiated ESCC cell EC109.KYSE150 or EC109 cells were transfected with negative control siRNA(si-NC group)and siRNA targeting FOXO3A(siFOXO3A group),respectively.CCK-8 assay was used to detect the proliferative capacity after 72 hour transfection,clone formation assay was used to evaluate clone formation capacity.Transwell chamber assay was used to detect migration and invasion,Western blot was used to detect the expressions of E-cadherin,N-cadherin,and Vimentin,and immunofluorescence was used to detect the aggregation of LC3B and p62 in cells after 24 hour transfection.Results:The mRNA and protein expressions of FOXO3A in the 2 ESCC cells were higher than those in HEEC(P<0.05).Compared with the si-NC group,the proliferative activity,the number of clone formation,the number of migrating cells,and the number of invasive cells in the siFOXO3A group were lower(P<0.05);the expression of N-cadherin and Vimentin decreased,and the expression of E-cadherin increased(P<0.05);the aggregation degree of p62 was weakened,and the aggregation degree of LC3B was enhanced(P<0.05).Conclusion:Knockdowning FOXO3A could inhibit the proliferation,migration,and invasion of ESCC cells,enhance autophagy level,and exert a tumor suppression effect.
作者
陈双双
杨莹
郑钊洋
张曼
陈希贤
肖楠
刘红春
CHEN Shuangshuang;YANG Ying;ZHENG Zhaoyang;ZHANG Man;CHEN Xixian;XIAO Nan;LIU Hongchun(The Second Clinical Medical College,Henan University of Traditional Chinese Medicine,Zhengzhou 450002;Department of Clinical Laboratory,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2024年第5期659-663,共5页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省自然科学基金项目(202300410464)
河南省医学科技攻关计划省部共建重点项目(SBGJ202002079)。
关键词
食管鳞状细胞癌
FOXO3A
增殖
迁移
侵袭
自噬
esophageal squamous cell carcinoma
FOXO3A
proliferation
migration
invasion
autophagy
